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Item type:Publication, Access status: without files , Automated Laser-Transfer Synthesis of High-Density Microarrays for Infectious Disease Screening(2022-06-01); ; ; Laser-induced forward transfer (LIFT) is a rapid laser-patterning technique for high-throughput combinatorial synthesis directly on glass slides. A lack of automation and precision limits LIFT applications to simple proof-of-concept syntheses of fewer than 100 compounds. Here, an automated synthesis instrument is reported that combines laser transfer and robotics for parallel synthesis in a microarray format with up to 10 000 individual reactions cm<sup>−</sup><sup>2</sup>. An optimized pipeline for amide bond formation is the basis for preparing complex peptide microarrays with thousands of different sequences in high yield with high reproducibility. The resulting peptide arrays are of higher quality than commercial peptide arrays. More than 4800 15-residue peptides resembling the entire Ebola virus proteome on a microarray are synthesized to study the antibody response of an Ebola virus infection survivor. Known and unknown epitopes that serve now as a basis for Ebola diagnostic development are identified. The versatility and precision of the synthesizer is demonstrated by in situ synthesis of fluorescent molecules via Schiff base reaction and multi-step patterning of precisely definable amounts of fluorophores. This automated laser transfer synthesis approach opens new avenues for high-throughput chemical synthesis and biological screening.Publicationtype: Journal Article8 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Assessing Polymer-Surface Adhesion with a Polymer Collection(2022-02-22); ; ; 11 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Combinatorial Synthesis of Peptoid Arrays via Laser-Based Stacking of Multiple Polymer Nanolayers(2019-03-01); 8 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , High-density peptide arrays help to identify linear immunogenic B-cell epitopes in individuals naturally exposed to malaria infection(2019-01-01); ; ; ; High-density peptide arrays are an excellent means to profile anti-plasmodial antibody responses. Different protein intrinsic epitopes can be distinguished, and additional insights are gained, when compared with assays involving the full-length protein. Distinct reactivities to specific epitopes within one protein may explain differences in published results, regarding immunity or susceptibility to malaria. We pursued three approaches to find specific epitopes within important plasmodial proteins, (1) twelve leading vaccine candidates were mapped as overlapping 15-mer peptides, (2) a bioinformatical approach served to predict immunogenic malaria epitopes which were subsequently validated in the assay, and (3) randomly selected peptides from the malaria proteome were screened as a control. Several peptide array replicas were prepared, employing particle-based laser printing, and were used to screen 27 serum samples from a malaria-endemic area in Burkina Faso, West Africa. The immunological status of the individuals was classified as "protected" or "unprotected" based on clinical symptoms, parasite density, and age. The vaccine candidate screening approach resulted in significant hits in all twelve proteins and allowed us (1) to verify many known immunogenic structures, (2) to map B-cell epitopes across the entire sequence of each antigen and (3) to uncover novel immunogenic epitopes. Predicting immunogenic regions in the proteome of the human malaria parasite Plasmodium falciparum, via the bioinformatics approach and subsequent array screening, confirmed known immunogenic sequences, such as in the leading malaria vaccine candidate CSP and discovered immunogenic epitopes derived from hypothetical or unknown proteins.Publicationtype: Journal Article13 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Combinatorial Synthesis of Macromolecular Arrays by Microchannel Cantilever Spotting (µCS)(2018-08-02); ; Surface-bound microarrays of multiple oligo- and macromolecules (e.g., peptides, DNA) offer versatile options in biomedical applications like drug screening, DNA analysis, or medical diagnostics. Combinatorial syntheses of these molecules in situ can save significant resources in regard to processing time and material use. Furthermore, high feature densities are needed to enable high-throughput and low sample volumes as generally regarded in combinatorial chemistry. Here, a scanning-probe-lithography-based approach for the combinatorial in situ synthesis of macromolecules is presented in microarray format. Feature sizes below 40 µm allow for the creation of high-density arrays with feature densities of 62 500 features per cm<sup>2</sup>. To demonstrate feasibility of this approach for biomedical applications, a multiplexed array of functional protein tags (HA- and FLAG-tag) is synthesized, and selective binding of respective epitope recognizing antibodies is shown. This approach uses only small amounts of base chemicals for synthesis and can be further parallelized, therefore, opening up a route to flexible, highly dense, and cost-effective microarrays.Publicationtype: Journal Article3 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , A Trifunctional Linker for Purified 3D Assembled Peptide Structure Arrays(2018-02-01); ; ; 5 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Antibody fingerprints in lyme disease deciphered with high density peptide arrays(2017-10-01); ; ; 6 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Identification of a Tetanus Toxin Specific Epitope in Single Amino Acid Resolution(2017-10-01); ; ; ; Vaccinations are among the most potent tools to fight infectious diseases. However, cross-reactions are an ongoing problem and there is an urgent need to fully understand the mechanisms of the immune response. For the development of a methodological workflow, the linear epitopes in the immune response to the tetanus toxin is investigated in sera of 19 vaccinated Europeans applying epitope mapping with peptide arrays. The most prominent epitope, appearing in nine different sera (923IHLVNNESSEVIVHK937), is investigated in a substitution analysis to identify the amino acids that are crucial for the binding of the corresponding antibody species − the antibody fingerprint. The antibody fingerprints of different individuals are compared and found to be strongly conserved (929ExxEVIVxK937), which is astonishing considering the randomness of their development. Additionally, the corresponding antibody species is isolated from one serum with batch chromatography using the amino acid sequence of the identified epitope and the tetanus specificity of the isolated antibody is verified by ELISA. Studying antibody fingerprints with peptide arrays should be transferable to any kind of humoral immune response toward protein antigens. Furthermore, antibody fingerprints have shown to be highly disease-specific and, therefore, can be employed as reliable biomarkers enabling the study of cross-reacting antigens.Publicationtype: Journal Article6 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Biochemical synthesis: Combinatorial chemistry in high-density array format(2016-09-01); ; Nesterov-Müller, AlexanderWe synthesize peptide arrays by using laser pulses to transfer amino acids onto a synthesis slide that are embedded in a polymer. After patterning these razor-thin spots, the coupling starts by heating the polymer that now serves as a solvent to couple monomers to the surface. The main advantages of this nanolayer solid phase chemistry are the high degree of automation, the very high spot density, and the in situ activation of monomers due to mixing different reagents in the stacked nanolayers.Publicationtype: Journal Article4 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Solid-material-based coupling efficiency analyzed with time-of-flight secondary ion mass spectrometry(2016)5